Screening and Characterization of Proteolytic Thermophiles Bacteria from Moinit Coastal Hot-Spring, North Sulawesi

Thermophilic bacteria have been recognized as the ideal producers of thermostable enzymes and therefore they have a long time become an interesting target for research. Thermophilic proteolytic bacteria from Moinit coastal hot-spring North Sulawesi Indonesia have been isolated and some of them, have been identified and reported. In this research, we reported another eleven thermophilic proteolytic bacteria from Moinit coastal hot-spring, North Sulawesi, Indonesia which designated as 1a, 1b, 2a, 3a, 3b, 4a, 11a, 11b, 16a, 16b, and 16c. Those thermophilic proteolytic bacteria had different colony and cell morphology characteristics, and biochemical characteristics. Bacterial isolate 1a, 1b, 2a, 16a and 16c were Gram negative-bacilli, 3a, 3b and 11b were Gram positive-bacilli, 4a, 11a and 16b were Gram positive-coccus. These bacterial isolates could produce proteolytic activities at 55oC. This showed their ability to produce thermostable protease extracellularly which has the potential to explore biotechnological industries.Keywords: Moinit, protease, screening, thermophilic  ABSTRAKBakteri termofilik dikenal sebagai produsen enzim termostabil dan menjadi target yang menarik untuk diteliti. Bakteri proteolitik termofil dari sumber air panas pantai Moinit Sulawesi Utara telah berhasil diisolasi dan bahkan 5 isolat bakteri telah diidentifikasi dan dilaporkan. Dalam tulisan ini, diuraikan sebelas isolat bakteri proteolitik termofil lainnya dalam kemampuannya menghasilkan protease dan karakteristik bakteri tersebut. Kesebelas bakteri adalah 1a, 1b, 2a, 3a, 3b, 4a, 11a, 11b, 16b, 16b, 16b, dan 16c yang menunjukkan karakteristik morfologi koloni dan sel bakteri, serta karakteristik biokimia yang berbeda. Isolat 1a, 1b, 2a, 16a, 16c adalah Gram negatif-basil, isolat 3a, 3b, 11b adalah Gram positif-basil, dan isolat 4a, 11a,16b adalah Gram positif-kokus. Keseluruhan isolat bakteri ini memiliki kemampuan aktivitas proteolitik pada 55oC. Hal ini menunjukkan kemampuan bakteri tersebut dalam menghasilkan protease termostabil ekstraseluler yang berpotensi untuk dieksplorasi dalam pemanfaatannya dalam bidang industri bioteknologi.Kata kunci: Moinit, protease, penapisan, termofili

Isolation of Symbiotic Bacteria with Red Algae from Tongkaina Waters, North Sulawesi

This study aims to isolate symbiotic bacteria with red algae from the waters of Tongkaina, North Sulawesi. The red algae taken were red algae that are similar to the genus Portieria and Gracilaria. Bacteria were cultured and isolated using Nutrient Agar (NA) + sea water. Before bacteria from red algae samples were cultured, each sample of red algae was crushed, homogenized and diluted. The results of this study, 5 bacterial isolates were successfully isolated from red algae similar to Portieria sp. and 5 bacterial isolates from red algae similar to Gracilaria sp. Each bacterial isolate has different morphological characteristics such as shape, colour, elevation, and edges. Of the 10 bacterial isolates, 3 bacterial cells are gram-positive cocci (round), and 7 of them are gram-negative bacilli (rod).Keywords: Red algae, bacteria, isolation, symbionts ABSTRAKTujuan dari penelitian ini adalah untuk mengisolasi bakteri simbion dengan alga merah dari perairan Tongkaina, Sulawesi Utara. Alga merah yang diambil adalah alga merah yang mirip dengan genus Portieria dan Gracilaria. Bakteri ditumbuhkan dan diisolasi menggunakan media Nutrien Agar (NA) + air laut.  Sebelum bakteri dari sampel alga merah ditumbuhkan, masing-masing sampel alga merah digerus, dihomogeniasi dan diencerkan.  Hasil penelitian ini, 5 isolat bakteri berhasil diisolasi dari alga merah mirip Portieria sp. dan 5 isolat bakteri dari alga merah mirip Gracilaria sp. Masing-masing isolat bakteri memiliki karakteristik morfologi yang berbeda seperti bentuk, warna, elevasi, dan tepian. Dari 10 isolat  bakteri  tersebut, 3 sel bakteri bersifat gram positif dengan bentuk bulat, dan 7 diantaranya bersifat gram negatif dengan bentuk batang.Kata kunci:  Alga merah, bakteri, isolasi, simbio

Analysis of Bacteria Community in the sediment from Bangka Island, North Sulawesi

Marine sediments are nutrient-rich and is a suitable habitat of bacteria. This research is a preliminary study of molecular analysis to identify the bacteria in the sediments from the littoral area that covered by mangroves in Bangka Island, North Sulawesi. The purposes of this study are to obtain the uncultivated bacterial DNA genome  which is used to identify the bacteria  and  bacterial community  in the sediments. Isolation of DNA genome from uncultured bacterial was carried out by following the genomic DNA extraction procedure using the DNeasy® PowerSoil Extraction Kit. Before isolating the bacterial DNA, sample were went through freezing and thawing processes. The DNA isolation result was subsequently tested using electrophoresis and UV-Vis spectrophotometers. Subsequently the genomic DNA was amplified using Polymerase Chain Reaction and the bacteria were identified using Next Generation Sequencing (NGS) analysis. The results of this study showed that the DNA of uncultured bacteria from sediment have the purity of 1.05 and the DNA amplification band was detected at 1300-1600bp. The bacteria in Bangka Island, North Sulawesi sediments were consisted of Gemmatimonadetes, Acidobacteria, Chioroflexi, Firmicutes, Bacteroidetes, Actinobacteria, Cyanobacteria and Proteobacteria respectively. Phylum Proteobacteria was found has the highest relative abundance  in the sediment.Keywords : Bacteria, Deoxyribo Nucleic Acid, Sediment, Uncultured. ABSTRAKSedimen laut merupakan suatu habitat yang kaya akan nutrient dan merupakan habitat dari bakteri. Penelitian ini merupakan tahapan awal dalam rangkaian analisis molekuler bakteri yang hidup di sedimen dari daerah litoral yang ditumbuhi oleh mangrove pada Pulau Bangka Sulawesi Utara. Tujuan penelitian ini adalah untuk mendapatkan DNA genom bakteri tanpa kultivasi yang digunakan dalam analisis jenis dan komunitas bakteri pada sedimen. Isolasi DNA genom bakteri tanpa kultivasi dilakukan dengan mengikuti prosedur Kit ekstraksi DNA DNeasy® PowerSoil. Sebelum tahap isolasi DNA bakteri, sampel diperlakukan proses freezing and thawing. Hasil isolasi DNA diuji menggunakan elektroforesis dan spektrofotometer UV-Vis. DNA genom diamplifikasi menggunakan Polymerase Chain Reaction dan ditentukan jenis bakteri dengan menggunakan Next Generation Sequencing analysis. Hasil penelitian menujukkan bahwa DNA bakteri tanpa kultivasi  memiliki kemurnian 1,05. DNA amplifikasi terdeteksi pada posisi 1300-1600bp. Dan jenis bakteri yang hidup pada sedimen di Pulau Bangka Sulawesi Utara, terdiri dari filum Gemmatimonadetes, Acidobacteria, Chioroflexi, Firmicutes, Bacteroidetes, Actinobacteria, Cyanobacteria dan Proteobacteria. Kelimpahan tertinggi bakteri yang hidup pada sedimen tersebut adalah  filum Proteobacteria.Kata kunci : Bakteri, Deoxyribo Nucleic Acid, Sedimen, Tanpa kultivasi

Amplification Of Bacterial Isolate Sf1 Associated With Sponge Facaplysynopsis sp. From Tongkeina, North Sulawesi

This study was conducted with the aim of amplifying the isolate of SF1 symbiont sponge Facaplysynopsis sp.  from Tongkeina, North Sulawesi. Samples are obtained and stored in the Lab. Molecular Biology and Marine Pharmacology, FPIK Unsrat. The genomic DNA of the samples was isolated using protocols from the Innu PREP Mini DNA Kit. The DNA of the SF1 symbionary bacteria was amplified by PCR (Polymerase Chain Reaction) using an 8F primer (5'-AGAGTITGATCCTGGCTA-3 ') and 1492 R (5'TACCTTACGACTT-3'). DNA bacteria SF1 successfully amplified marked by the appearance of the band of DNA that looks less clear, with a length of 600 bp.Keywords: Bacterial Isolate, Sponge,  AmplificationABSTRAKPenelitian ini dilakukan dengan tujuan mengamplifikasi isolat bakteri SF1 simbion spons Facaplysynopsis sp dari perairan Tongkeina, Sulawesi Utara. Sampel diperoleh dan tersimpan di Lab. Biologi Molekuler dan Farmasitika Laut, FPIK Unsrat. DNA genom dari sampel diisolasi menggunakan protokol dari Innu PREP DNA Mini Kit. DNA bakteri simbion SF1 diamplifikasi dengan PCR (Polymerase Chain Reaction) dengan menggunakan primer 8F (5’-AGAGTITGATCCTGGCTA-3’) dan 1492 R (5’TACCTTACGACTT-3’). DNA bakteri SF1 berhasil diamplifikasi ditandai dengan munculnya pita DNA yang terlihat kurang jelas, dengan panjang 600 bp.Kata Kunci: Bakteri Simbion, Spons, Amplifikas

ISOLASI BAKTERI LAUT DARI PERAIRAN MALALAYANG, SULAWESI UTARA

Marine bacteria have a lot of potential in exploring the enzyme that can be developed, such as a producer of proteorhodopsin, act as hydrocarbon chlorlastic and can degrade oil. This study aims to obtain isolates and can characterize the bacterial morphology. Malalayang Waters is one of the marine bacterial habitats that has potential area to be studied. This study aims to isolate marine bacteria from Malalayang Waters. These marine bacteria first were diluted into sea water before they were grown on Nutrient Agar (NA). Based on the results of this study it was found that marine bacterial isolated were separated based on their morphological characteristics. The dominant morphological characteristics were yellow whites which dominant shape were irregular.Keywords: bacterial, dilution, isolation.  Bakteri laut memiliki banyak potensi yang dapat dikembangkan. Seperti penghasil proteorhodopsin, berperan sebagai hidrokarbonoklastik dan dapat mendegradasi minyak. Perairan Malalayang merupakan salah satu habitat bakteri laut yang belum diteliti. Penelitian ini bertujuan untuk mengisolasi bakteri laut dari Perairan Malalayang. Bakteri laut ditumbuhkan pada media agar + air laut, selain itu bakteri juga dilakukan pengenceran terhadap air laut sebelum bakteri ditumbuhkan pada media Nutrient Agar (NA). Berdasarkan hasil penelitian ini isolat bakteri  laut ditemukan, bakteri tersebut dipisahkan berdasarkan karakteristik morfologinya. Karakteristik morfologi yang dimiliki dominan berwarna putih kuning dan memiliki bentuk yang dominan tidak teratur. Hal ini dapat memperlihatkan perbedaan bakteri laut dari Perairan Malalayang yang tumbuh.Kata kunci: bakteri, pengenceran, isolasi

STRUKTUR KOMUNITAS HUTAN MANGROVE DI DESA BUDO KECAMATAN WORI KABUPATEN MINAHASA UTARA

Mangrove forest can grow on the muddy soil, along the coast and around the river estuaries that are affected by the tidal. The mangrove forest has a specific vegetation structure. Study on the structure of the mangrove forest community, was carried out in the Budo Village. Mangrove community structure data was taken using line transect method. A 100 m line was established from the sea perpendicular to the coast. A total of three line transects was made and each line has five 10 x 10 m plots with 20 m distance between plots. Distance between transect is 50m. This study found that three mangrove species Rhizophora apiculata, Sonneratia alba, and Bruguiera gymmorrhiza were found in the three transects during this study. The highest species biodiversity index was found in transect two (H’ = 0.97). R. apiculata has the highest density and relative density 0.05 ind/m2 and 67.57%. Both R. apiculata and S. alba have the highest frequency and relative frequency of 1.00 and 50% respectively. The highest species coverage and relative coverage area is belonging to S. alba in transect two, 25,89 and 63.57%. S. alba has the highest Important value index 140.72 in transect two.Keywords: Budo Village, Mangrove Forest, Community Structure.

Isolation and Antibacterial Activity assay of Endophytic Symbiont Bacteria on Seaweed Gracilaria verrucosa originated from Batu Meja Tongkaina Beach, North Sulawesi

Seaweed can produce bioactive compounds such as antibacterial. Seaweed co-exists with bacteria endophytes. Entophytic bacteria are bacteria that live in host tissues and have the ability to protect the host itself by producing antibacterial compounds against pathogens. Therefore, the endophytic bacteria of seaweed symbionts can be utilized to produce antibacterial compounds. Bacteria can be mass-cultured because of their fast-growing characteristic. The aim of this study was to determine the antibacterial activity from endophytic symbiont bacteria on seaweed Gracilaria verrucosa originated from Batu Meja Tongkaina Beach, North Sulawesi. Isolation of bacteria was carried out by dilution steps from 101 to 10־3 times, then cultivation on Nutrient Agar media to obtain a single colony of the bacteria. Antibacterial activity was tested used paper disc diffusion method. The pathogens used were bacterial strains of Escherichia coli, Staphylococcus aureus, Streptococcus mutans, and Salmonella typhi. Moreover, antibiotics were used as positive controls. The antibacterial ability of the symbiotic bacteria was measured based on inhibition zones around the paper disc containing the isolates of endophytic bacteria. A total of 6 endophytic bacterial isolates were found, namely isolate B, C, D, E, F and G, characterized by different morphological features. The results showed that isolates B and C having inhibition zone of 0.5 – 1.0 mm against S. typhi, indicating that these two isolates produce antibacterial compounds with a weak ability  against S. typhi

Isolation and Screening the Symbiont Bacteria of the Sponge Dragmacidon sp from Manado Bay, North Sulawesi that Producing Chitinase and Protease

Enzymes are important in the technology industry and hydrolytic enzymes, such as chitinase and protease are commonly used for it. Various types of microorganisms such as bacteria can produce hydrolytic enzymes.  Sponge-associated bacteria are excellent sources of extracellular hydrolytic enzymes because the surface and internal spaces of sponges are richer in nutrients. The aim of this study was to isolate and screen the bacteria of the sponge Dragmacidon sp symbiotic from Manado Bay, North Sulawesi that producing chitinase and protease   Symbiont bacteria were grown in Zobell 1226 E medium with a dilution of 10-4. Bacterial isolation was carried out based on the morphological characteristics of the colony. Chitinase and protease activity was carried out by growing each bacterial isolate in chitin and protein media at 36oC for 48 hours. Chitinase and protease activities were indicated by the formation of a clear zone around the bacterial colony, however, the clear zone for chitinase activity was observed after pouring the Lugol's solution. Based on this study, 8 isolates bacteria of the symbiotic spongy Dragmacidon sp from Manado Bay, North Sulawesi were isolated based on morphological characteristics. The colony of the bacteria is generally white with an irregular shape. Four isolates, namely 1, 2, 3, and 8 had chitinase activity with chitinolytic indexes were 1.7; 1.5; 1.4, and 1.3, respectively. Six isolates, namely 1, 2, 3, 4, 5, and 6 had protease activity with proteolytic indexes were 1.4; 1.8; 3.1; 1.3; 1.8; and 2.5, respectively.Keywords: Bacteria; Chitinolytic; Proteolytic; Symbiont; SpongeAbstrakEnzim menempati posisi penting dalam bidang teknologi dan industri. Enzim yang banyak digunakan dalam bidang industri adalah enzim hidrolase. Enzim dapat diisolasi dari berbagai jenis mikroorganisme seperti bakteri. Bakteri yang berasosiasi dengan spons merupakan sumber enzim hidrolitik ekstraseluler yang sangat baik karena permukaan dan ruang internal spons lebih kaya nutrisi. Penelitian ini bertujuan untuk mengisolasi dan menguji aktivitas kitinase dan protease bakteri simbion spons Dragmacidon sp dari Teluk Manado. Bakteri simbion spons ditumbuhkan dalam media Zobell 1226 E pada pengenceran 10-4. Isolasi bakteri dilaksanakan berdasakan karakteristik morfologi. Aktivitas kitinase dan protease dilaksanakan dengan menumbuhkan setiap isolat bakteri dalam media kitin dan protein pada suhu 36oC selama 48 jam. Aktivitas kitinase dan protease ditandai dengan terbentuknya zona bening di sekitar koloni bakteri yang mana untuk kitinase diamati setelah diberi larutan lugol. Berdasarkan penelitian ini, 8 isolat bakeri simbion spon Dragmacidon sp dari Teluk Manado, Sulawesi Utara berhasil diisolasi berdasarkan karakteristik morfologi. Isolat bakteri umumnya berwarna putih dengan bentuk ireguller. Empat isolat yakni 1, 2, 3, dan 8 memiliki aktivitas kitinase dan enam isolat yakni 1, 2, 3, 4, 5, dan 6 yang memiliki aktivitas protease. Indeks kitinolitik dari masing-masing keempat isolat bakteri secara berturut turut adalah 1,7; 1,5; 1,4; dan 1,3 dengan kategori bernilai rendah dan indeks proteolitik adalah 1,4; 1,8; 3,1; 1,3; 1,8; dan 2,5 dengan kategori bernilai rendah sampai tinggi.Kata kunci: Bakteri; Kitinolitik; Proteolitik; Simbion; Spon

OKSIGEN TERLARUT DAN pH DI AIR SISIPAN SEDIMEN MANGROVE DAN PESISIR DI DESA BULUTUI KECAMATAN LIKUPANG BARAT

This research was conducted in Bulutui Village, West Likupang District, related to theproblems raised how is the solubility of Dissolved Oxygen (DO) and the pH of sediment-insertedwater at mangrove locations, and on the coast, then this research aims to find out how the solubility ofoxygen (DO), and acidity (pH) of sediment-inserted water in mangrove ecosystems and on the coastin Bulutui Village, West Likupang District. Based on the results of measurements that have beencarried out in Bulutui Village, West Likupang District, it was found that the value of the dissolvedoxygen content at the mangrove location was in the range of 0.36 – 1.98 ppm, with an average of0.98 ppm, the pH value ranged from 6.16 – 6.89 with an average value of 6.04. In coastal locations,the dissolved oxygen content ranges from 1.26 to 2.87 ppm, with an average of 2.09 ppm, the pHvalue ranges from 6.78 to 8.90, with an average value of 8.34. The statistical t test showed that therewas a difference between the dissolved oxygen values in the sedimentary water in the mangrovesand the coast, as well as the acidity (pH). Keywords: sediment; Inset Water, Dissolved Oxygen, pH Acidity ABSTRAK Penelitian ini dilakukan di Desa Bulutui Kecamatan Likupang Barat, berkaitan denganpermasalahan yang dikemukakan bagaimanakah kelarutan Oksigen Terlarut (DO) dan pH air sisipansedimen pada lokasi mangrove, dan di Pesisir pantai, selanjutnya penelitian ini bertujuan untukmengetahui bagaimana kelarutan oksigen (DO), dan keasaman (pH) air sisipan sedimen padaekosistem mangrove dan di pesisir di Desa Bulutui Kecamatan Likupang Barat. Berdasarkan hasilpengukuran yang telah dilakukan di Desa Bulutui Kecamatan Likupang Barat ditemukan nilai darikandungan oksigen terlarut pada lokasi mangrove adalah berkisar 0,36 – 1,98 ppm, dengan rataratanya 0,98 ppm, nilai pH berkisar 6,16 – 6,89 dengan nilai rata rata 6,04. Pada Lokasi pesisirdengan kandungan oksigen terlarut berkisar 1,26 – 2,87 ppm, dengan rata rata 2,09 ppm nilai pHberkisar 6,78 – 8,90, dengan nilai rata – rata 8,34. Uji statistika t test menunjukkan adanyaperbedaan antara nilai oksigen terlarut pada air sisipan sedimen di mangrove, dan pesisir, demikianjuga keasaman (pH). Kata Kunci: Sedimen; Air sisipan, Oksigen Terlarut, Keasaman p

Community structure and distribution of Gastropoda in the intertidal area of the Waleo districts coast north Minahasa

This research is to know community structure and gastropod distribution include species density (individual/m2), relative density (%), diversity indices (H’), Evenness indices (E), Richness indices (R), Dominance indices (D) and Dispersion indices (Id).This inestigation was carried out February 2020 in intertidal area, Waleo beach in Minahasa Northern. Quadrat transect placed via area sampling based on substrate difference. Placing quadrat transect was centr on three points that different with distance with point 100 m and distance between quadran 10 m with size quadrat 50 cm2. Based on this research was find 22 species include Cypraea tigris, Terebra areolata, Cymbiola nivosa oblita, Euplica borealis, Turbo argyrostomus, Cypraea moneta, Conus cumingii, Cypraea errones, Tectus fenestrats, Nerita ascensionis, Nerita exuvi, Nerita albicilla, Nerita polita, Nerita Picea, Vasum turbinellus, Lunella cinerea, Nassarius globosus, Nassarius pullus, Vexillum acupictum, Clypeomorus pellucida, Vexillum ebenus, Terebralia sulcata. Five order include Neogastropoda, Littorinimorpha, Cycloneritida, Caenogastropoda, Trochida. Analysis result to species density is the highest 0,40 (individual/m2),relative density, Nerita polita 12,63 %, index of diversity,H’= 3,…, index of evenness, E = 0,97, index of richness, R = 0,22 and index of dominance, range D =0,13-0,18 concluded that no species dominantly. Gastropoda distribution pattern in intertidal area based 3 stations that showed all pattern of clumed distribution.   Keywords: Gastropoda, Intertidal, density, spread pattern   Abstrak Penelitian ini bertujuan untuk mengetahui struktur komunitas dan pola penyebaran gastropoda meliputi Kepadatan spesies (Ind/m2), Kepadatan Relatif (%),Indeks Keanekaragaman (H’), Indeks Keseragaman (E), Indeks Kekayaan spesies (R), Indeks Dominansi (D), dan indeks Sebaran. Penelitian ini di lakukan pada bulan februari 2020 yang berlokasi di daerah intertidal pantai Waleo Kabupaten Minahasa Utara. Penempatan transek kuadran di tentukan melalui teknik area sampling berdasarkan perbedaan substrat, penempatan transek dan kuadran di pusatkan di tiga titik yang berbeda-beda dengan jarak antara titik 100m dan jarak antara kuadran 10m dengan ukuran kuadran 50x50cm. Dari hasil penelitiaan di temukan 22 spesis meliputi Cypraea tigris, Terebra areolata, Cymbiola nivosa oblita, Euplica borealis, Turbo argyrostomus, Cypraea moneta, Conus cumingii, Cypraea errones, Tectus fenestrats, Nerita ascensionis, Nerita exuvi, Nerita albicilla, Nerita polita, Nerita Picea, Vasum turbinellus, Lunella cinerea, Nassarius globosus, Nassarius pullus, Vexillum acupictum, Clypeomorus pellucida, Vexillum ebenus, Terebralia sulcata. Dan 5 ordo Neogastropoda, Littorinimorpha, Cycloneritida, Caenogastropoda, Trochida. Hasil analisis terhadap kepadatan spesis nilai tertinggi terdapat pada spesis N.polita sebesar 0,40 Ind/m² dan kepadatan realtif tertinggi juga terdapat pada spesis N.polita sebesar 12,63 %, nilai indeka keanekaragaman H’= 9,49 yang merupakan kategori tinggi, nilai dari indeks keseragaman secara keseluruhan  (E)=0,97 dalam kondisi , dan nilai indeks kekayaan spesis yaitu 0,22,  indeks Dominansi (D) = 0,13 – 0,18 disimpulkan tidak ada yang mendominasi, Pola sebaran gastropoda di daerah intertidal dari ketiga stasiun menunjukan pola sebaran secara mengelompok. Wilayah pantai Waleo Kabupaten Minahasa Utara memiliki suhu 31,3ºC. Derajat keasaman pH yang di peroleh yaitu 7 Salinitas yang di peroleh sebesar 34‰ yang masih dalam kisaran baik untuk gastropoda. Kata kunci : Gastropoda, intertidal, kepadatan, pola sebara